FLT3-HDAC double-target inhibitor as well as preparation method and application thereof

Abstract

The invention relates to an FLT3-HDAC double-target inhibitor and a preparation method and application thereof.The FLT3-HDAC double-target inhibitor is of a structure shown in the formula I. The FLT3-HDAC double-target inhibitor not only shows good FLT3 and HDAC enzyme inhibition activity, but also has certain anti-proliferative activity on solid tumors and hematoma in vitro, especially on acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia, acute and a strong anti-tumor effect is shown. As a double-target antitumor drug based on FLT3 and HDACs, the compound has further research value. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a FLT3-HDAC dual-target inhibitor and a preparation method and application thereof, belonging to the technical field of medicine. Background Art

[0002] Acute myeloid leukemia (AML) is caused by abnormal proliferation, cloning, and differentiation of cells in the hematopoietic system, and is characterized by genetic heterogeneity. Although the treatment strategies for AML have improved significantly, from intensive chemotherapy (cytarabine + anthracyclines) to targeted small molecule drugs, the treatment prospects for AML remain bleak due to the problems of drug resistance and disease relapse. Among them, FMS-like tyrosine kinase 3 (FLT3) mutations are closely associated with poor clinical outcomes and the development of treatment resistance, especially in patients with relapsed / refractory AML (R / R AML). On this basis, the combination of FLT3 inhibitors and other anti-AML drugs is a commonly used strategy to address drug resistance, improve efficacy, and improve the overall survival (OS) of AML patients, such as double therapy and triple therapy. The former is represented by gilteritinib and 7+3 induction chemotherapy (NCT02236013), gilteritinib and azacitidine (NCT02752035), and gilteritinib and venetola (NCT03625505); the latter includes the combination of gilteritinib, azacitidine and venetola (NCT05520567), and the combination of quizartini, decitabine and venetola (NCT03661307). In recent years, related studies have reported that synergistic inhibition of FLT3 and histone deacetylase (HDAC) can enhance the therapeutic response of AML cells with FLT3 mutations and reduce drug resistance. These results indicate that dual inhibition of FLT3 and HDAC is expected to be an effective strategy for the treatment of FLT3-driven AML.

[0003] FLT3 is a receptor tyrosine kinase (RTK) that plays a key role in regulating multiple signal transduction cascades, including the Janus kinase (JAK) / signal transducer activator of transcription 5 (STAT5), phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT), and rat sarcoma (RAS) / mitogen-activated protein kinase (MAPK) pathways. Patients with AML harboring FLT3 mutations are more likely to relapse compared with patients harboring wild-type FLT3 (FLT3-WT). Approximately 25% of adult AML patients harbor FLT3 internal tandem duplication (FLT3-ITD) mutations, and 10% harbor FLT3 tyrosine kinase domain (FLT3-TKD) mutations. Notably, treatment of FLT3-mutated R / R AML typically involves the use of FLT3 inhibitors such as midoshuin, quizartinib, and gilteritinib alone or in combination with other inhibitors. On the other hand, HDAC has the ability to remove lysine acetyl groups from histone or non-histone substrates and is subdivided into four branches based on intracellular localization and functional mechanisms, including class I (HDAC1, 2, 3, 8), class IIa (HDAC 4, 5, 7, 9), class IIb (HDAC 6, 10), class III (Sirtuin 1-7) and class IV (HDAC11). The catalytic activity of class I, IIa and IV HDAC members depends on zinc ions, while the catalytic activity of class III HDAC members depends on NAD + To date, five HDAC inhibitors have been approved by the FDA for clinical use in the treatment of hematological malignancies, including cedabenamide (CS055), romidepsin (FK 228), voritastat (SAHA), belinostat (PDX101), and panobinostat (LBH-589).

[0004] Although the combination of FLT3 and HDAC inhibitors has certain advantages in the treatment of AML, potential adverse drug-drug interactions limit the realization of synergy, such as complex dosing regimens and negative effects on pharmacokinetics. In contrast, FLT3 / HDAC dual-target inhibitors can circumvent these potential disadvantages. However, the FLT3 / HDAC dual-target inhibitors reported so far have no significant therapeutic advantages over single FLT3 inhibitors.

[0005] Therefore, the development of an efficient FLT3-HDAC dual-target inhibitor has important clinical significance. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides a FLT3-HDAC dual-target inhibitor and a preparation method and application thereof. The present invention is achieved through the following technical solutions:

[0007] A FLT3-HDAC dual-target inhibitor or a pharmaceutically acceptable salt or a stereoisomer thereof, characterized in that the FLT3-HDAC dual-target inhibitor has a structure shown in Formula I:

[0008]

[0009] In formula I, R1 is hydroxyl or 2-aminophenyl,

[0010] X is an aromatic heterocycle or one of the following linking groups:

[0011]

[0012] A is one of the following structures:

[0013]

[0014] R2 is a hydrogen atom or one of the following structures

[0015]

[0016] Preferably according to the present invention, in the FLT3-HDAC dual-target inhibitor formula I,

[0017] R1 is hydroxyl or 2-aminophenyl;

[0018] X is one of the following linking groups:

[0019]

[0020] A is piperazinyl or piperidin-4-ylpiperazinyl;

[0021] R2 is tetrahydropyranylamino.

[0022] The FLT3-HDAC dual-target inhibitor is selected from one of the following compounds:

[0023] 6-ethyl-3-((4-(4-(4-(5-(hydroxyamino)-5-oxopentanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11a);

[0024] 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxohexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11b);

[0025] 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxoheptanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11c);

[0026] 6-ethyl-3-((4-(4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11d);

[0027] 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11e);

[0028] 6-ethyl-3-((4-(4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11f);

[0029] 6-ethyl-3-((4-(4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11 g);

[0030] 6-ethyl-3-((4-(4-(4-(4-(hydroxycarbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11h);

[0031] (E)-6-ethyl-3-((4-(4-(4-(4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11i);

[0032] (E)-6-ethyl-3-((4-(4-(4-(4-((4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)carbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11j);

[0033] 2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)N-hydroxy-pyrimidine-5-carboxamide (14);

[0034] 3-((4-(4-(4-(4-((2-aminophenyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (17a);

[0035] N-(2-aminophenyl)-2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pyrimidine-5-carboxamide (17b);

[0036] 6-ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25a);

[0037] 6-ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25b);

[0038] 6-ethyl-3-((4-(4-(3-(hydroxyamino)-3-oxopropyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25c);

[0039] 6-ethyl-3-((4-(4-(4-(hydroxyamino)-4-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25d);

[0040] 6-ethyl-3-((4-(4-(5-(hydroxyamino)-5-oxopentyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25e);

[0041] 6-ethyl-3-((4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25f);

[0042] 6-ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25 g);

[0043] 6-ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25h);

[0044] (E)-6-ethyl-3-((4-(4-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25i);

[0045] (E)-6-ethyl-3-((4-(4-(4-((4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)amino)-2-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25j);

[0046] (E)-6-ethyl-3-((4-(4-(2-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25k);

[0047] (E)-6-ethyl-3-((4-(4-(3-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)propyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (251);

[0048] (E)-6-ethyl-3-((4-(4-(4-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)butyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25m);

[0049] 2-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25n);

[0050] 2-((3-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)propyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (25o);

[0051] 2-(4-((4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25p).

[0052] 2-(4-((4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25q)

[0053] The second object of the present invention is to provide a method for preparing a FLT3-HDAC dual-target inhibitor, comprising the following steps:

[0054] Compound 1 reacts with 4-aminotetrahydropyran to obtain compound 2. Meanwhile, compounds 3 and 18 react with 1-fluoro-2-methoxy-4-nitrobenzene to obtain compounds 4 and 19. Compounds 4 and 19 are further reduced to compounds 5 and 20. Compounds 2 and 5 or 20 are subjected to Buchwald-Hartwig coupling reaction to obtain compounds 6 and 21. Compounds 6 and 21 are subjected to oxidation reaction and removal of Boc protecting groups to obtain compounds 8 and 23. Compounds 8 and 23 are further reacted with 9a-u, 12, 15a and 15b to generate 10a-j, 13, 16a, 16b and 24a-q, wherein the ester groups of compounds 10a-j and 24a-q can be converted into hydroxamic acid groups in an alkaline hydroxylamine solution to obtain target compounds 11a-j and 25a-q. Compounds 13, 16a and 16b are subjected to removal of protecting groups in an acidic environment to obtain target compounds 14, 17a and 17b.

[0055] A method for preparing a FLT3-HDAC dual-target inhibitor, the preparation method comprising any of the following methods:

[0056] The preparation methods of compounds 11a-11j are as follows:

[0057]

[0058] Reagents and conditions: (a) 4-aminotetrahydropyran, triethylamine, acetonitrile, 80°C, 6 hours; (b) 1-fluoro-2-methoxy-4-nitrobenzene, potassium carbonate, N,N-dimethylformamide, 100°C, 8 hours; (c) 10% Pd / C, tetrahydrofuran, room temperature, 24 hours; (d) palladium acetate, 4,5-bis(diphenylphosphino-9,9-dimethylxanthene), cesium carbonate, 1,4-dioxane, 100°C, 12 hours; (e) 30% hydrogen peroxide, potassium carbonate, dimethyl sulfoxide, room temperature, 12 hours; (f) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (g) 1) 9a-d, EDCI, DMAP, room temperature, 8 hours; 2) 9e-j, potassium carbonate, acetonitrile, 80°C, 6 hours; (h) NH2OH, KOH, anhydrous methanol, room temperature, 4 hours;

[0059] The preparation methods of compounds 14, 17a and 17b are as follows:

[0060]

[0061] Reagents and conditions: (a) 12, triethylamine, acetonitrile, 80°C, 8 hours; (b) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (c) 15a and 15b, triethylamine, acetonitrile, 80°C, 8 hours; (d) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours;

[0062] The preparation method of compound 25a-q is as follows:

[0063]

[0064] Reagents and conditions: (a) 1-fluoro-2-methoxy-4-nitrobenzene, potassium carbonate, N,N-dimethylformamide, 100°C, 8 hours; (b) 10% Pd / C, tetrahydrofuran, room temperature, 24 hours; (c) 2, palladium acetate, 4,5-bis(diphenylphosphino-9,9-dimethylxanthene), cesium carbonate, 1,4-dioxane, 100°C, 12 hours; (d) 30% hydrogen peroxide, potassium carbonate, dimethyl sulfoxide, room temperature, 12 hours; (e) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (f) 1) 9c and 9d, EDCI, DMAP, room temperature, 8 hours; 2) 9e-u, potassium carbonate, acetonitrile, 80°C, 6 hours; (g) NH2OH, KOH, anhydrous methanol, room temperature, 4 hours.

[0065] A pharmaceutical composition contains a therapeutically effective amount of the FLT3-HDAC dual-target inhibitor and pharmaceutically acceptable excipients.

[0066] The FLT3-HDAC dual-target inhibitor is used in the preparation of drugs for treating tumor diseases related to the target.

[0067] The tumors are lung cancer, liver cancer, non-small cell lung cancer, colon cancer, skin cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, lymphoma, esophageal cancer, gastrointestinal cancer, nasopharyngeal cancer, leukemia, and myeloma.

[0068] Beneficial Effects

[0069] 1. The compounds of the present invention not only exhibit good FLT3 and HDAC enzyme inhibitory activity, can inhibit FLT3 and HDAC activity in cells, downregulate ERK phosphorylation levels, but also have strong in vitro anti-tumor activity.

[0070] 2. The present invention takes the potent FLT3 inhibitor gilteritinib as the research basis, evaluates the effects of gilteritinib, SAHA and their combined use on the vitality of AML cells carrying FLT3 mutations, and successfully develops a more effective FLT3 / HDAC dual-target inhibitor, which is the first reported dual-target anti-tumor drug based on DNMT1 and HDAC, and its tumor inhibition effect is significantly better than the combined use of gilteritinib and SAHA. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 In vivo antitumor activity of MOLM-13 xenografted BALB / c nude mice;

[0072] Among them, A is the change in mouse tumor volume during the treatment period, B is the volume of mouse tumor after the treatment, and C is the weight of mouse tumor after the treatment. DETAILED DESCRIPTION

[0073] The reagents, raw materials or intermediates used in the present invention are all commercially available or can be prepared according to literature methods. The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The present invention is further described below in conjunction with the examples, but is not limited thereto.

[0074] Example 1: Preparation of Intermediate 8

[0075] 3-Bromo-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2)

[0076] 4-Aminotetrahydropyran (1.64 g, 16.23 mmol) was added to a solution of compound 1 (4 g, 16.23 mmol) and triethylamine (4.92 g, 48.69 mmol) in acetonitrile (50 mL) and reacted at 80 °C for 6 hours. Then, the solvent was removed by rotary evaporation under reduced pressure and the residue was washed with 100 mL of water, followed by extraction of the aqueous phase with ethyl acetate (30 mL × 3). The organic phase was separated and dried over anhydrous Na2SO4, and then concentrated to obtain a crude product. The latter was purified by silica gel column chromatography to obtain compound 2 with a yield of 89.4%. 1 H NMR (400MHz, DMSO) δ7.67 (d, J=7.7Hz, 1H), 4.18–4.05 (m, 1H), 3.94–3.86 (m, 2H), 3.46–3.37 (m,2H),2.64(q,J=7.3Hz,2H),1.82–1.74(m,2H),1.69–1.58(m,2H),1.17(t,J=7.3Hz,3H).

[0077] Tert-butyl-4-(1-(2-methoxy-4-nitrophenyl)piperidin-4-yl)piperazine-1-carboxylate (4)

[0078] 1-Fluoro-2-methoxy-4-nitrobenzene (2.1 g, 10.23 mmol) was added to a DMF solution (25 mL) of compound 3 (2.76 g, 10.23 mmol) and potassium carbonate (2.83 g, 20.45 mmol) and reacted at 100°C for 8 hours. Then, the reaction solution was diluted with 250 mL of water, and the aqueous phase was extracted with ethyl acetate (80 mL × 3). The organic phase was separated and dried over anhydrous Na2SO4, and then concentrated to obtain a crude product. The latter was purified by silica gel column chromatography to obtain compound 4 with a yield of 92.8%. The synthesis method of compound 19 is the same as the operation steps. 1 H NMR (400MHz, DMSO) δ7.96(s,1H),7.82(dd,J=8.9,2.6Hz,1H),7.68(d,J=2.6Hz,1H),7.01(d,J=8.9Hz,1H),3.91(s,3H),3.71(d, J=12.0Hz,2H),3.30(d,J=5.4Hz,2H),2.74(s,4H),2.46(t,J=5.0Hz,5H),1.83(d,J=12.1Hz,2H),1.60–1.52(m,2H),1.40(s,9H).

[0079] Tert-butyl-4-(1-(4-amino-2-methoxyphenyl)piperidin-4-yl)piperazine-1-carboxylate (5)

[0080] 10% Pd / C was added to the THF solution of compound 4, and the mixture was reacted at room temperature for 24 hours under a hydrogen atmosphere. The reaction solution was then filtered through diatomaceous earth, and the filtrate was collected and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 5 with a yield of 91.7%. The synthesis method of compound 20 was the same as the above operation steps. 1 H NMR (400MHz, DMSO) δ7.95(s,1H),6.60(d,J=8.4Hz,1H),6.21(d,J=2.3Hz,1H),6.05(dd,J=8.3,2.3Hz,1H),4.67(s,2H), 3.67(s,3H),3.29(s,2H),3.14(d,J=11.0Hz,2H),2.46–2.41(m,5H),1.77–1.69(m,2H),1.56–1.47(m,2H),1.39(s,9H).

[0081] Tert-butyl-4-(1-(4-((3-cyano-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazine-1-carboxylate (6)

[0082] Compound 2 was added to an anhydrous 1,4-dioxane solution (50 mL) of compound 5, palladium acetate, (9,9-dimethyl-9H-oxanthene-4,5-diyl)bis(diphenylphosphine) and cesium carbonate, and heated to 100°C for 12 hours under nitrogen protection. Then, the reaction solution was diluted with 200 mL of water, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phase was separated and dried over anhydrous Na2SO4, and then concentrated to obtain a crude product. The latter was purified by silica gel column chromatography to obtain compound 6 with a yield of 64.3%. The synthesis method of compound 21 is the same as the operation steps. 1 H NMR (400MHz, DMSO) δ8.60(s,1H),7.15(dd,J=8.5,,2.3Hz,1H),3.75(s,2H),3.24(s,2H),2.60–2.52(m,7H),2.45(d, J=5.4Hz,3H),2.33(s,1H),1.76(d,J=12.0Hz,4H),1.57(dd,J=16.0,7.9Hz,4H),1.40(s,9H),1.12(t,J=7.4Hz,3H).

[0083] Tert-butyl-4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazine-1-carboxylate (7)

[0084] In a dimethyl sulfoxide solution of compound 6 (1g, 1.61mmol) and potassium carbonate (555mg, 4.03mmol), 30% hydrogen peroxide (1g, 3.21mmol) was slowly added dropwise, and the mixture was reacted at room temperature for 12 hours. Then the mixture was diluted with 150mL of water and the aqueous phase was extracted with ethyl acetate (40mL×3). The organic phase was dried over anhydrous magnesium sulfate and concentrated to obtain a crude product, which was then purified by silica gel column chromatography to obtain compound 7. The synthesis method of compound 21 is the same as the operation steps. Bright yellow solid, yield 52.4%. 1 HNMR (400MHz, DMSO) δ11.02 (s, 1H), 7.53 (s, 1H), 7.30–7.20 (m, 2H), 7.02 (d, J = 2. 3Hz,1H),6.80(dd,J=15.9,8.1Hz,2H),4.10(s,1H),3.93(d,J=11.1Hz,2H),3.81 (s,3H),3.39(s,2H),2.62–2.54(m,6H),2.46(d,J=5.5Hz,4H),2.33(s,1H),1.82 (dd,J=24.7,12.3Hz,4H),1.65–1.57(m,4H),1.40(s,9H),1.19(t,J=7.3Hz,3H).

[0085] 4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazine-1-ammonium salt (8)

[0086] Compound 7 (498 mg, 0.78 mmol) was added to 4M HCl / EtOAc solution and stirred at room temperature for 12 hours. Subsequently, the reaction solution was filtered and the filter cake was recrystallized from ethyl acetate to obtain compound 8. The synthesis method of compound 21 was the same as the above operation steps. Yellow solid, yield 88.9%. 1H NMR(400MHz,DMSO)δ11.46(s,1H),10.16(s,2H),9.83(s,2H),7.62(s,2H),7.50– 7.33(m,1H),7.24(d,J=1.9Hz,1H),6.98(s,1H),4.13(s,1H),4.00(s,3H),3.94(d d,J=12.2,8.3Hz,2H),3.68(s,4H),3.55(s,4H),3.38(dd,J=12.7,10.6Hz,2H),2 .62(q,J=7.3Hz,2H),1.92–1.82(m,2H),1.75–1.65(m,2H),1.21(t,J=7.3Hz,3H).

[0087] Example 2: Preparation of Targets 11a-j, 25a-i

[0088] 5-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)-5-pentanoic acid methyl ester (10a)

[0089] Compound 9a (49 mg, 0.33 mmol) was added to a dichloromethane solution (15 mL) of compound 8 (171 mg, 0.28 mmol), EDCI (69 mg, 0.36 mmol) and DMAP (103 mg, 0.84 mmol). After addition, the mixture was reacted at room temperature for 8 hours. After completion, the reaction solution was washed with saturated sodium bicarbonate solution and saturated sodium chloride solution in sequence, the organic phase was separated, dried over anhydrous magnesium sulfate, and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 10a. The synthesis methods of compounds 10b-d, 24a and 24b were the same as the above operation steps. Yellow solid, yield 91.4%. 1 H NMR (400MHz, DMSO) δ11.02 (s, 1H), 7.54 (d, J = 3.3Hz, 1H), 7.30–7.21 (m, 2H), 7.0 2(d,J=2.4Hz,1H),6.81(t,J=8.7Hz,2H),4.12(s,1H),3.94(s,2H),3.81(s,3H), 3.59(s,3H),3.44–3.39(m,6H),2.58(d,J=7.4Hz,2H),2.45(d,J=5.3Hz,2H),2. 34(t,J=7.5Hz,4H),1.85–1.76(m,4H),1.71–1.51(m,6H),1.19(t,J=7.4Hz,3H).

[0090] 6-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)-6-hexanoic acid methyl ester (10b)

[0091] Yellow solid, yield 84.2%. 1 H NMR (400MHz, DMSO) δ11.02 (s, 1H), 7.53 (d, J = 3.2Hz, 1H), 7.33–7.20 (m, 2H), 7.02 (d,J=2.3Hz,1H),6.85–6.76(m,2H),4.11(s,1H),3.92(d,J=11.3Hz,2H),3.81(s ,3H),3.58(s,3H),3.47–3.37(m,6H),2.59(t,J=7.2Hz,2H),2.47–2.42(m,2H),2 .32(t,J=7.2Hz,4H),1.86–1.75(m,4H),1.69–1.47(m,8H),1.19(t,J=7.3Hz,3H).

[0092] 7-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)-6-heptanoic acid ethyl ester (10c)

[0093] Yellow solid, yield 73.5%. 1 H NMR (400MHz, DMSO) δ11.02(s,1H),7.53(s,1H),7.30–7.21(m,2H),7.02(d,J=2.3Hz,1H),6.85–6.76(m,2H),4.04(q,J=7.1Hz,4H),3.92(d,J=10.9 Hz,2H),3.81(s,3H),3.42(s,6H),2.57(t,J=7.4Hz,2H),2.46(s,2H),2.3 7–2.28(m,4H),1.85–1.73(m,4H),1.69–1.43(m,10H),1.25–1.16(m,5H).

[0094] 8-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)-6-octanoic acid methyl ester (10d)

[0095] Yellow solid, yield 71.2%.1 H NMR (400MHz, DMSO) δ11.02(s,1H),7.53(s,1H),7.30–7.22(m,2H),7.02(d,J=2.3Hz, 1H),6.81(t,J=8.8Hz,2H),4.11(d,J=7.4Hz,1H),3.92(d,J=11.1Hz,2H),3.81(s,3H) ,3.58(s,3H),3.42(s,6H),2.61–2.56(m,2H),2.29(dd,J=8.5,6.2Hz,4H),1.87–1.76 (m,4H),1.66–1.55(m,4H),1.50(q,J=8.1Hz,6H),1.26(s,2H),1.19(t,J=7.3Hz,3H).

[0096] 6-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)hexanoic acid methyl ester (10e)

[0097] Compound 9e (194 mg, 0.93 mmol) was added to a solution of compound 8 (281 mg, 0.46 mmol) and triethylamine (85 mg, 0.84 mmol) in acetonitrile. After addition, the mixture was reacted at 80°C for 8 hours. The solvent was removed by evaporation under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 10e. The synthesis methods of compounds 10f-j, 13, 16a, 16b, 24c-i, 27a-c, 30a-f, 36a-c, 41 and 43 were the same as the above operation steps.

[0098] Yellow solid, yield 74.4%. 1 H NMR (400MHz, DMSO) δ11.02(s,1H),7.53(s,1H),7.30–7.21(m,2H),7.02(d,J=2.3Hz,1H) ,6.81(t,J=8.4Hz,2H),4.11(s,1H),3.92(d,J=10.6Hz,2H),3.81(s,3H),3.58(s,3H),3 .39(s,2H),2.61–2.57(m,2H),2.37–2.27(m,6H),1.85(d,J=14.1Hz,4H),1.63(d,J=8.3 Hz,2H),1.52(q,J=7.7Hz,4H),1.41(s,2H),1.25(d,J=8.4Hz,2H),1.19(t,J=7.4Hz,3H).

[0099] 7-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)heptanoic acid methyl ester (10f)

[0100] Yellow solid, yield 83%. 1 H NMR (400MHz, DMSO) δ11.02(s,1H),7.53(s,1H),7.30–7.20(m,2H),7.02(d,J=2.3Hz ,1H),6.80(dd,J=13.1,8.1Hz,2H),4.11(d,J=7.5Hz,1H),3.92(d,J=11.0Hz,2H),3 .81(s,3H),3.58(s,3H),3.37(d,J=9.8Hz,2H),2.58(q,J=7.4Hz,8H),2.38–2.22(m ,6H),1.85(d,J=13.5Hz,4H),1.70–1.46(m,8H),1.27(s,4H),1.19(t,J=7.3Hz,3H).

[0101] 8-(4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)octanoic acid methyl ester (10 g)

[0102] Yellow solid, yield 67.7%. 1 H NMR (400MHz, DMSO) δ11.03 (s, 1H), 7.54 (d, J = 3.1Hz, 1H), 7.31–7.21 (m, 2H), 7.03 (d,J=2.3Hz,1H),6.82(t,J=8.3Hz,2H),3.93(d,J=11.1Hz,2H),3.81(s,3H),3.58 (s,3H),3.39(s,2H),2.59(s,2H),2.29(dd,J=9.4,5.4Hz,6H),1.85(d,J=12.7Hz, 4H),1.69–1.60(m,4H),1.53(d,J=7.2Hz,4H),1.27(s,6H),1.19(t,J=7.3Hz,3H).

[0103] Methyl 4-((4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)benzoate (10h)

[0104] Yellow solid, yield 73.1%. 1 H NMR (400MHz, DMSO) δ7.95(d,J=7.9Hz,2H),7.56–7.45(m,3H),7.32–7.21(m,2H),6.82(dd,J=13.0,8.1Hz,2H),4.11(q,J=5.3Hz,2H),3.9 2(d,J=11.2Hz,2H),3.85(s,2H),3.81(s,2H),3.38(s,2H),2.59(q,J=7.3Hz,2H),1.85(d,J=13.1Hz,4H),1.67–1.59(m,4H),1.20(s,3H).

[0105] (E)-methyl 3-(4-((4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)phenyl)acrylate (10i)

[0106] Yellow solid, yield 75.8%. 1 H NMR(400MHz,DMSO)δ11.04(s,1H),7.74–7.62(m,3H),7.54(s,1H),7.38(s,2H),7 .31–7.20(m,2H),7.03(d,J=2.3Hz,1H),6.81(dd,J=12.4,8.1Hz,2H),6.64(d,J=1 5.9Hz,1H),4.12(s,1H),3.93(s,2H),3.81(s,3H),3.73(s,3H),3.60(s,2H),3.3 8(s,2H),2.59(t,J=7.4Hz,4H),1.85(d,J=13.3Hz,4H),1.65(s,4H),1.20(s,3H).

[0107] (E)-methyl 3-(4-((4-((4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)benzamido)methyl)phenyl)acrylate (10j)

[0108] Yellow solid, yield 50.9%. 1H NMR (400MHz, DMSO) δ11.04(s,1H),9.09(s,1H),7.88(d,J=7.8Hz,2H),7.73–7.61(m,4H),7.55(s,1H), 7.43(s,2H),7.36(d,J=8.0Hz,2H),7.31–7.22(m,2H),7.03(d,J=2.2Hz,1H),6.82(t,J=8.1Hz,2H),6.6 2(d,J=16.1Hz,1H),4.50(d,J=5.9Hz,2H),4.12(q,J=5.2Hz,2H),3.92(d,J=9.0Hz,2H),3.81(s,3H),3 .72(s,3H),3.39(s,2H),2.58(d,J=7.4Hz,4H),1.85(d,J=12.7Hz,4H),1.73–1.55(m,4H),1.18(s,3H).

[0109] 6-Ethyl-3-((4-(4-(4-(5-(hydroxyamino)-5-oxopentanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11a)

[0110] Potassium hydroxide (3.3 g, 50 mmol) was added to anhydrous methanol solution containing hydroxylamine hydrochloride (2.35 g, 33.5 mmol), and the reaction was continued at 0°C for 0.5 hours. The reaction solution was then filtered, the filtrate was collected, and the filtrate was added dropwise to anhydrous methanol solution containing compound 10a (166 mg, 0.25 mmol), and the reaction was continued at room temperature for 4 hours. After the reaction was completed, the solvent was removed by evaporation under reduced pressure, the residue was dissolved in 5 mL of water, and the pH value was adjusted to about 7 with 1 M hydrochloric acid, a solid was precipitated, filtered, and the filter cake was collected. The latter was recrystallized from anhydrous ethanol to obtain compound 11a. The synthesis methods of compounds 11b-j, 25a-i, 28a-c, 31a-f, 37a-c and 42 were the same as the operation steps. Yellow solid, yield 63.8%, mp 190-192°C. 1H NMR (400MHz, DMSO) δ11.03(s,1H),10.37(s,1H),8.69(s,1H),7.54(d,J=3.3Hz,1H),7.33– 7.19(m,2H),7.02(d,J=2.3Hz,1H),6.81(t,J=7.3Hz,2H),4.17–4.05(m,1H),3.95–3.86(m ,2H),3.81(s,3H),3.46–3.38(m,4H),2.58(d,J=7.3Hz,2H),2.47–2.43(m,2H),2.33–2.26 (m,3H),1.99(t,J=7.3Hz,2H),1.88–1.76(m,4H),1.74–1.51(m,6H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.45,170.12,169.38,152.74,152.21,150.61,136.90,135.81,131.97,118.73,112.12,105.25,66.91 ,61.58,56.16,50.98,49.53,49.03,48.04,45.75,41.86,32.64,32.06,28.64,24.67,21.38,11.42.HRMS-ESI(m / z)calc.for C 33 H 49 N9O6[M+H] + 668.3833,found 668.3829.HPLC t R =6.80min (98.66% purity).

[0111] 6-Ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxohexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11b)

[0112] Yellow solid, yield 53.3%, mp 196-198°C. 1H NMR(400MHz,DMSO)δ11.03(s,1H),10.36(s,1H),8.70(s,1H),7.62–7.47(m,1H),7.35–7.21 (m,2H),7.02(d,J=2.3Hz,1H),6.81(t,J=7.9Hz,2H),4.18–4.06(m,1H),3.97–3.88(m,2H), 3.81(s,3H),3.42(q,J=8.5Hz,4H),2.58(q,J=7.3Hz,2H),2.47–2.42(m,2H),2.29(q,J=7.3 Hz,3H),1.95(t,J=6.8Hz,2H),1.90–1.74(m,4H),1.70–1.41(m,8H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.76,170.12,169.45,152.75,152.21,150.61,136.91,135.81,131.96,118.74,112.14,112.08,105.28,66.91,61 .58,56.17,50.98,49.58,49.07,48.04,45.84,41.87,32.65,32.59,32.40,28.66,25.36,24.86,24.66,11.42.HRMS-ESI(m / z)calc.forC 34 H 51 N9O6[M+H] + 682.3989,found 682.3985.HPLC t R =8.76min (97.28% purity).

[0113] 6-Ethyl-3-((4-(4-(6-(hydroxyamino)-6-oxoheptanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11c)

[0114] Yellow solid, yield 33.3%, mp 185-187°C. 1H NMR (400MHz, DMSO) δ11.03(s,1H),10.34(s,1H),8.68(s,1H),7.54(d,J=3.2Hz,1H),7.32–7 .20(m,2H),7.02(d,J=2.3Hz,1H),6.81(t,J=8.2Hz,2H),4.16–4.06(m,1H),3.92(d,J=10.9H z,2H),3.81(s,3H),3.47–3.38(m,4H),2.61–2.56(m,2H),2.46(d,J=5.6Hz,2H),2.29(q,J= 11.0Hz,3H),1.93(t,J=7.3Hz,2H),1.89–1.75(m,4H),1.69–1.42(m,9H),1.33–1.15(m,5H). 13 C NMR (101MHz, DMSO) δ170.82,170.12,169.52,152.74,152.21,150.61,136.90,135.81,131.95,118.72,112.12,112.06,105.25,66.91,61.57 ,56.16,50.98,49.57,49.06,48.04,45.83,41.85,32.68,32.65,32.60,28.85,28.65,25.48,25.02,24.67,11.42.HRMS-ESI(m / z)calc.forC 35 H 53 N9O6[M+H] + 696.4147,found696.4142.HPLC t R =8.14min (97.26% purity).

[0115] 6-Ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11d)

[0116] Yellow solid, yield 38.2%, mp 190-192°C. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.32(s,1H),8.64(s,1H),7.52(d,J=3.2Hz,1H),7.29–7.18(m,2H),7.03 (d,J=2.4Hz,1H),6.80(dd,J=17.8,8.1Hz,2H),4.17–4.06(m,1H),3.96–3.88(m,2H),3.81(s,3H),3.39(s,4 H),3.36(d,J=2.1Hz,2H),2.58(q,J=7.3Hz,2H),2.45(d,J=11.7Hz,2H),2.35–2.24(m,3H),1.93(t,J=7.4Hz ,2H),1.88–1.76(m,4H),1.66–1.58(m,9H),1.53–1.39(m,4H),1.25(d,J=6.8Hz,4H),1.19(t,J=7.4Hz,3H). 13 C NMR (101MHz, DMSO) δ170.82,170.12,169.52,152.74,152.21,150.61,136.90,135.81,131.95,118.72,112.12,112.06,105.25,66.91,61.5 7,56.16,50.98,49.57,49.06,48.04,45.83,41.85,32.68,32.65,32.60,28.85,28.65,25.48,25.02,24.67,11.42.HRMS-ESI(m / z)calc.for C 36 H 55 N9O6[M+H] + 710.4298,found 710.4293.HPLC t R =6.28min (95.77% purity).

[0117] 6-Ethyl-3-((4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11e)

[0118] Pale red solid, yield 22%, mp 198-200°C. 1H NMR (400MHz, DMSO) δ11.03(s,1H),10.35(s,1H),7.53(s,1H),7.34–7.15(m,2H),7.04(s,1H), 6.80(t,J=8.4Hz,2H),4.12(d,J=10.0Hz,1H),3.93(d,J=11.0Hz,2H),3.81(s,3H),3.40–3.33 (m,2H),2.59(q,J=7.4Hz,2H),2.46(d,J=10.0Hz,4H),2.34(s,2H),2.22(q,J=9.2Hz,4H),1.9 8–1.77(m,6H),1.69–1.60(m,2H),1.54–1.46(m,4H),1.38(q,J=7.5Hz,2H),1.25–1.16(m,5H). 13 CNMR(101MHz,DMSO)δ170.12,169.53,152.74,152.21,150.62,136.97,135.78,131.93,118.69,112.13,112.10,105.33,66.91,61.55,58.38,5 8.29,56.18,53.78,50.97,49.12,48.04,46.13,32.73,32.66,28.84,27.04,26.53,25.58,24.67,11.83,11.41,8.30.HRMS-ESI(m / z)calc.for C 34 H 53 N9O5[M+H] + 668.4198,found668.4193.HPLC t R =10.63min (96.46% purity).

[0119] 6-ethyl-3-((4-(4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11f)

[0120] Pale yellow solid, yield 30%, mp 183-185°C. 1H NMR (400MHz, DMSO) δ11.04(s,1H),10.42(s,1H),7.54(d,J=3.3Hz,1H),7.33–7.21(m,2H) ,7.03(d,J=2.3Hz,1H),6.83(dd,J=8.1,5.4Hz,2H),4.15–4.06(m,1H),3.92(dd,J=10.3, 4.1Hz,2H),3.82(s,3H),3.38(d,J=11.3Hz,2H),2.86(s,8H),2.64–2.56(m,4H),1.98–1. 82(m,6H),1.70–1.61(m,4H),1.57–1.45(m,4H),1.29–1.23(m,4H),1.19(t,J=6.6Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.53,152.72,152.21,150.59,136.55,135.95,132.02,118.78,112.10,111.99,105.13,66.91,61.65,60 .23,57.04,56.13,51.75,50.60,48.04,34.08,32.63,28.77,28.18,26.74,25.45,24.69,21.24,14.56,11.43.HRMS-ESI(m / z)calc.for C 35 H 55 N9O5[M+H] + 682.4356,found 682.4351.HPLC t R =8.62min (99.25% purity).

[0121] 6-ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11 g)

[0122] Pale yellow solid, yield 31.7%, mp 198-200°C. 1H NMR (400MHz, DMSO) δ11.03(s,1H),10.34(s,1H),8.75(s,1H),7.53(s,1H),7.34–7.16(m,2H),7.04(s ,1H),6.80(t,J=8.9Hz,2H),4.19–4.08(m,1H),3.93(d,J=11.0Hz,2H),3.81(s,3H),3.38(d,J=11.5H z,2H),2.59(q,J=7.4Hz,2H),2.46(d,J=11.3Hz,4H),2.33(s,2H),2.28–2.13(m,4H),1.94(t,J=7.3H z,2H),1.90–1.74(m,4H),1.64(d,J=14.2Hz,2H),1.58–1.44(m,4H),1.38(s,2H),1.28–1.16(m,9H). 13 C NMR (101MHz, DMSO) δ170.12,169.57,152.74,152.21,150.62,136.97,135. 78,131.91,118.68,112.14,112.09,105.32,66.91,61.55,60.22,58.48,5 6.18,53.80,50.98,49.15,48.04,32.74,32.66,29.17,29.05,28.85,27.3 6,26.79,25.57,24.67,21.23,14.56,11.41,8.36.HRMS-ESI(m / z)calc.for C 36 H 57 N9O5[M+H] + 696.4147,found 696.4142.HPLC t R =6.94min (99.58% purity).

[0123] 6-Ethyl-3-((4-(4-(4-(4-(hydroxycarbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11h)

[0124] Pale yellow solid, yield 35.3%, mp 218-220°C. 1H NMR (400MHz, DMSO) δ11.18(s,1H),11.02(s,1H),8.99(s,1H),7.72(d,J=7.7Hz,2H),7.52(s,1H ),7.38(d,J=7.7Hz,2H),7.26(d,J=8.6Hz,1H),7.23–7.18(m,1H),7.03(d,J=2.3Hz,1H),6.80(d d,J=16.2,8.1Hz,2H),4.12(s,1H),3.92(d,J=11.0Hz,2H),3.81(s,3H),3.53(s,2H),3.37(d,J= 11.5Hz,2H),2.59(q,J=7.4Hz,2H),2.00–1.74(m,4H),1.74–1.49(m,4H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.12,152.77,152.23,150.62,135.96,132.04,129.19,127.34,118.83,112.17,112.11 ,105.30,66.90,56.19,50.65,48.86,48.05,45.79,32.65,29.48,24.68,11.43,8.98.HRMS-ESI(m / z)calc.for C 36 H 49 N9O5[M+H] + 668.3881,found 668.3875.HPLC t R =7.48min (95.18% purity).

[0125] (E)-6-ethyl-3-((4-(4-(4-(4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11i)

[0126] Yellow solid, yield 30%, mp 220-222°C. 1H NMR (400MHz, DMSO) δ11.01(s,1H),10.76(s,1H),9.03(s,1H),7.51(d,J=7.7Hz,3H),7.33(d,J =7.8Hz,2H),7.28–7.18(m,2H),7.03(s,1H),6.79(dd,J=17.5,8.0Hz,2H),6.46(d,J=15.7Hz, 1H),4.11(d,J=9.3Hz,1H),3.92(d,J=10.0Hz,2H),3.80(s,3H),3.47(s,2H),2.63–2.52(m,6H ),2.39(s,2H),2.29(s,1H),1.83(t,J=14.0Hz,4H),1.71–1.51(m,4H),1.19(t,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO)δ170.13,163.23,152.72,152.21,150.61,140.15,138.54 ,136.82,135.84,134.06,131.96,129.84,127.82,119.13,118.71,112.11,1 12.07,105.25,66.91,62.09,61.65,60.23,56.15,53.11,50.85,48.98,48.0 4,32.64,28.57,24.68,21.22,14.55,11.62,11.40.HRMS-ESI(m / z)calc.for C 38 H 51 N9O5[M+H] - 714.4089, found 714.4093.HPLCt R =7.63min (97.72% purity).

[0127] (E)-6-ethyl-3-((4-(4-(4-(4-((4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)carbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11j)

[0128] Yellow solid, yield 38.5%, mp 205-206°C. 1H NMR (400MHz, DMSO) δ11.01(s,1H),9.03(t,J=6.1Hz,1H),7.86(d,J=7.9Hz,2H),7.55–7.44(m,3H),7.37(dd,J= 19.3,8.1Hz,4H),7.29–7.18(m,2H),7.02(d,J=2.3Hz,1H),6.79(dd,J=17.0,8.0Hz,2H),6.43(d,J=15.8Hz,1H) ,4.49(d,J=5.9Hz,2H),4.12(s,1H),3.92(d,J=10.8Hz,2H),3.80(s,3H),3.50(s,2H),3.40(d,J=7.7Hz,2H),2. 58(q,J=7.4Hz,4H),2.39(s,4H),2.27(s,1H),1.83(t,J=14.2Hz,4H),1.69–1.50(m,4H),1.19(t,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO)δ170.13,166.59,163.19,152.73,152.21,150.62,142.31,141.74, 138.45,136.93,135.79,133.86,133.39,131.95,129.11,128.19,127.94,127.68,119. 14,118.69,112.12,112.06,105.25,66.91,62.17,61.53,58.25,56.15,53.55,50.96, 49.05,48.04,45.98,42.86,32.65,28.77,24.69,11.42,8.23.HRMS-ESI(m / z)calc.for C 46 H 58 N 10 O5[M+H] + 847.4546,found 847.4542.HPLC t R =9.62min (99.97% purity).

[0129] Example 3: Preparation of Targets 14, 17a, and 17b

[0130] 2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)N-((tetrahydro-2H-pyran-2-yl)oxy)pyrimidine-5-carboxamide (13)

[0131] Yellow solid, yield 33.3%, mp 167-169°C. 1 H NMR(400MHz,DMSO)δ11.55(s,1H),11.03(s,1H),9.38(s,2H),8.69(s,1H),7.54 (s,1H),7.26(d,J=17.4Hz,2H),7.03(s,1H),6.87–6.77(m,2H),4.96(s,1H),4.1 2(s,2H),3.93(d,J=10.4Hz,2H),3.82(s,3H),3.53(d,J=11.8Hz,2H),2.59(d,J= 7.3Hz, 4H), 1.86 (d, J = 12.2Hz, 4H), 1.72 (s, 4H), 1.65–1.51 (m, 6H), 1.24 (s, 3H).

[0132] 2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)N-hydroxy-pyrimidine-5-carboxamide (14)

[0133] Compound 13 was added to 4M hydrochloric acid / ethyl acetate (10 mL) and reacted at room temperature for 12 hours. The reaction solution was filtered and the filter cake was recrystallized with ethanol to obtain compound 14. The synthesis method of compounds 17a, 17b and 44 was the same as the operation steps. Brown solid, yield 28.1%, mp 216-218°C. 1 H NMR (400MHz, DMSO) δ11.93(s,1H),11.32(d,J=48.7Hz,2H),8.78(s,2H),7.59(d,J=29.4Hz ,2H),7.36(s,1H),7.20(s,1H),6.94(d,J=7.5Hz,1H),4.84(d,J=13.7Hz,2H),4.16–4.10( m,1H),3.94(d,J=14.9Hz,5H),3.61(d,J=12.9Hz,8H),3.42–3.35(m,2H),2.62(q,J=7.4Hz ,2H),2.36(s,4H),1.87(dd,J=12.2,4.0Hz,2H),1.69–1.60(m,2H),1.20(t,J=7.3Hz,3H). 13C NMR(101MHz,DMSO)δ170.13,166.59,163.19,152.73,152.21,150.62,142.31,141.74, 138.45,136.93,135.79,133.86,133.39,131.95,129.11,128.19,127.94,127.68,119. 14,118.69,112.12,112.06,105.25,66.91,62.17,61.53,58.25,56.15,53.55,50.96, 49.05,48.04,45.98,42.86,32.65,28.77,24.69,11.42,8.23.HRMS-ESI(m / z)calc.for C 33 H 45 N 11 O5[MH] - 674.3503,found674.3498.HPLC t R =10.16min (99.32% purity).

[0134] tert-Butyl (2-(4-((4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)benzamido)phenyl)carbamate (16a)

[0135] Yellow solid, yield 70.7%. 1 H NMR (400MHz, DMSO) δ11.05(s,1H),9.87(s,1H),9.65(s,2H),8.70(s,1H),7.96(d,J=7.9Hz,2H),7.52 (dd,J=14.3,7.4Hz,4H),7.29(d,J=8.5Hz,1H),7.27–7.13(m,4H),7.04(s,1H),6.82(dd,J=14.5,8.0 Hz,2H),4.10(t,J=5.3Hz,1H),3.92(d,J=11.3Hz,2H),3.82(s,3H),3.68(s,2H),3.50(s,2H),3.39(s ,4H),2.62–2.53(m,6H),2.14(s,2H),1.85(d,J=12.8Hz,4H),1.64(s,4H),1.45(s,9H),1.23(s,3H).

[0136] Tert-butyl (2-(4-((4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)pyrimidine-5-carbamoyl)phenyl)carbamate (16b)

[0137] Yellow solid, yield 77.2%. 1 H NMR (400MHz, DMSO) δ11.04(s,1H),9.67(s,1H),8.94(s,2H),8.62(s,1H),7.61(d,J=8.1 Hz,1H),7.54(s,1H),7.50–7.46(m,1H),7.31–7.17(m,4H),7.12(t,J=7.7Hz,2H),7.03( s,1H),6.83(dd,J=14.8,8.2Hz,2H),4.13–4.10(m,1H),3.92(d,J=9.2Hz,2H),3.82(s,3 H),2.59(d,J=7.4Hz,4H),1.85(d,J=12.7Hz,4H),1.63(s,4H),1.45(s,9H),1.23(s,3H).

[0138] 3-((4-(4-(4-((2-aminophenyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (17a)

[0139] Yellow solid, yield 28.3%, mp 174-176°C. 1 H NMR (400MHz, DMSO) δ11.42(s,1H),10.69(s,1H),8.23(d,J=8.0Hz,2H),7.87(d,J=7.9Hz,2H),7.68 –7.54(m,3H),7.51(d,J=7.8Hz,1H),7.45–7.33(m,3H),7.32–7.13(m,2H),6.96(d,J=7.5Hz,1H),4 .55(s,2H),4.12(s,1H),3.95(d,J=20.4Hz,5H),3.73(s,8H),3.43–3.33(m,2H),2.62(q,J=7.4Hz, 2H), 2.34 (d, J=8.5Hz, 4H), 1.86 (dd, J=12.4, 4.1Hz, 2H), 1.73–1.61 (m, 2H), 1.19 (d, J=7.3Hz, 3H). 13C NMR(101MHz,DMSO)δ170.12,165.62,152.75,152.21,150.61,143.61,135 .90,133.86,131.98,129.15,128.23,127.15,126.91,123.86,118.78,116 .72,116.61,112.14,112.06,105.24,66.91,61.77,56.16,50.76,48.96, 48.04,45.98,32.65,28.43,26.82,24.68,11.42.HRMS-ESI(m / z)calc.for C 42 H 54 N 10 O4[M+H] + 763.4349,found763.4345.HPLC t R =5.67min (97.79% purity).

[0140] N-(2-aminophenyl)-2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pyrimidine-5-carboxamide (17b)

[0141] Yellow solid, yield 27%, mp 238-240°C. 1 H NMR (400MHz, DMSO) δ11.02 (d, J = 14.3Hz, 1H), 9.55 (s, 1H), 8.92 (s, 2H), 7.54 (s, 1H) ),7.35–7.18(m,2H),7.17–7.09(m,1H),7.08–6.90(m,2H),6.90–6.68(m,3H),6.58 (t,J=8.1Hz,1H),4.96(s,2H),4.11(s,1H),3.95–3.75(m,7H),3.35(q,J=11.2Hz, 8H),2.68–2.55(m,4H),1.85(d,J=13.4Hz,4H),1.62(s,4H),1.18(t,J=7.6Hz,3H). 13C NMR (101MHz, DMSO) δ170.12,163.11,161.77,158.62,152.76,152.22,150.61,143.78,131.99,127.36,127.02,123.27,118.80,116.53,11 6.37,112.15,112.07,105.25,66.91,56.17,50.81,48.89,48.05,45.99,32.65,28.35,24.68,11.42,11.31,9.01.HRMS-ESI(m / z)calc.for C 39 H 50 N 12 O4[M+H] + 763.4349,found763.4345.HPLC t R =10.97min (99.93% purity).

[0142] Tert-Butyl-4-(2-methoxy-4-nitrophenyl)piperazine-1-carboxylate (19)

[0143] Yellow solid, yield 87.3%. 1 H NMR (400MHz, DMSO) δ7.84 (dd, J=8.8, 2.6Hz, 1H), 7.70 (d, J=2.6Hz, 1H), 7.03 (d, J= 8.9Hz,1H),3.91(s,3H),3.46(t,J=5.0Hz,4H),3.15(d,J=5.4Hz,4H),1.42(s,9H).

[0144] Tert-butyl-4-(4-amino-2-methoxyphenyl)piperazine-1-carboxylate (20)

[0145] Pale yellow solid, yield 88.1%. 1 H NMR (400MHz, DMSO) δ6.61(d,J=8.3Hz,1H),6.23(d,J=2.3Hz,1H),6.06(dd,J=8.3,2.4 Hz,1H),4.76(s,2H),3.68(s,3H),3.42–3.37(m,5H),2.76–2.70(m,5H),1.41(s,9H).

[0146] Tert-butyl-4-(4-((3-cyano-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazine-1-carboxylate (21)

[0147] Yellow solid, yield 41.5%. 1 H NMR (400MHz, DMSO) δ8.67 (s, 1H), 7.20 (dd, J = 8.6, 2.3Hz, 1H), 7.14–7.05 (m, 2 H),6.80(d,J=8.6Hz,1H),3.94(s,1H),3.92–3.84(m,2H),3.76(s,3H),3.45( s,4H),3.25(t,J=11.9,2H),2.94–2.78(m,4H),2.56(q,J=7.3Hz,2H),1.76(d d,J=12.2,4.1Hz,2H),1.70–1.61(m,2H),1.42(s,9H),1.13(t,J=7.3Hz,3H).

[0148] Tert-butyl-4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazine-1-carboxylate (22)

[0149] Yellow solid, yield 57.5%. 1 H NMR (400MHz, DMSO) δ11.06 (s, 1H), 7.54 (d, J = 3.5Hz, 1H), 7.32 (dd, J = 8.6, 2.3Hz, 1H), 7. 24(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.82(dd,J=14.7,8.0Hz,2H),4.12(d,J=8.0H z,1H),3.92(d,J=9.8Hz,2H),3.82(s,3H),3.45(s,4H),3.40(s,2H),2.94–2.81(m,4H), 2.58(d,J=7.4Hz,2H),1.85(d,J=12.6Hz,2H),1.71–1.54(m,2H),1.19(t,J=7.3Hz,3H).

[0150] 4-(4-((3-Carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazine-1-ammonium salt (23)

[0151] Bright yellow solid, yield 85.5%. 1H NMR (400MHz, DMSO) δ11.18(s,1H),9.42(s,2H),7.14–7.05(m,2H),6.87(s,1H),4.13(s,1H),3.96–3.90(m,2H), 3.87(s,3H),3.49–3.32(m,5H),2.63–2.57(m,2H),1.90–1.81(m,2H),1.72–1.60(m,2H),1.20(t,J=7.4Hz,3H).

[0152] 7-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)-7-heptanoic acid methyl ester (24a)

[0153] Yellow solid, yield 78.1%. 1 H NMR (400MHz, DMSO) δ11.99(s,1H),11.06(s,1H),7.55(s,1H),7.32(dd,J=8.5,2.7Hz,1H),7.25(s,1H),7 .03(d,J=2.7Hz,1H),6.86–6.78(m,2H),4.15–4.09(m,1H),3.92(d,J=11.6Hz,2H),3.83(d,J=2.1Hz,3H), 3.58(s,3H),2.87(d,J=22.9Hz,4H),2.62–2.55(m,2H),2.37–2.25(m,6H),2.19(td,J=7.4,2.2Hz,2H),1 .86(d,J=12.7Hz,2H),1.70–1.60(m,2H),1.50(p,J=6.9Hz,7H),1.32–1.25(m,6H),1.20(t,J=7.4Hz,3H).

[0154] 8-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)-8-octanoic acid methyl ester (24b)

[0155] Yellow solid, yield 75%. 1H NMR (400MHz, DMSO) δ11.99(s,1H),11.06(s,1H),7.55(s,1H),7.32(dd,J=8.5,2.7Hz,1H),7.25(s,1H),7 .03(d,J=2.7Hz,1H),6.86–6.78(m,2H),4.15–4.09(m,1H),3.92(d,J=11.6Hz,2H),3.83(d,J=2.1Hz,3H), 3.58(s,3H),2.87(d,J=22.9Hz,4H),2.62–2.55(m,2H),2.37–2.25(m,6H),2.19(td,J=7.4,2.2Hz,2H),1 .86(d,J=12.7Hz,2H),1.70–1.60(m,2H),1.50(p,J=6.9Hz,7H),1.32–1.25(m,6H),1.20(t,J=7.4Hz,3H).

[0156] Ethyl 3-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)propanoate (24c)

[0157] Yellow solid, yield 81.3%. 1 H NMR (400MHz, DMSO) δ11.00 (s, 1H), 7.54 (d, J = 3.2Hz, 1H), 7.27 (dd, J = 8.5, 2.3Hz, 1H),7.22(d,J=3.3Hz,1H),7.03(d,J=2.3Hz,1H),6.81(dd,J=8.1,5.9Hz,2H),4.0 7(q,J=7.1Hz,4H),3.92(d,J=11.3Hz,2H),3.80(s,3H),2.90(s,4H),2.60(dd,J=1 3.3, 6.3Hz, 4H), 1.85 (d, J = 12.3Hz, 2H), 1.68–1.58 (m, 2H), 1.19 (t, J = 7.0Hz, 6H).

[0158] Methyl 4-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)butanoate (24d)

[0159] Yellow solid, yield 83.6%. 1H NMR(400MHz,DMSO)δ11.00(s,1H),7.54(d,J=3.2Hz,1H),7.27(dd,J=8.5,2.3Hz,1H), 7.22(d,J=3.3Hz,1H),7.03(d,J=2.3Hz,1H),6.81(dd,J=8.1,5.9Hz,2H),4.07(q,J=7 .1Hz,4H),3.92(d,J=11.3Hz,2H),3.80(s,3H),2.90(s,4H),2.60(dd,J=13.3,6.3Hz, 4H), 1.85 (d, J=12.3Hz, 2H), 1.63 (qd, J=12.1, 4.5Hz, 2H), 1.19 (dd, J=7.6, 6.4Hz, 6H).

[0160] 5-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)pentanoic acid methyl ester (24e)

[0161] Yellow solid, yield 82.2%. 1 H NMR(400MHz,DMSO)δ11.00(s,1H),7.54(d,J=3.2Hz,1H),7.27(dd,J=8.5,2.3Hz,1H), 7.22(d,J=3.3Hz,1H),7.03(d,J=2.3Hz,1H),6.81(dd,J=8.1,5.9Hz,2H),4.07(q,J=7 .1Hz,4H),3.92(d,J=11.3Hz,2H),3.80(s,3H),2.90(s,4H),2.60(dd,J=13.3,6.3Hz, 4H), 1.85 (d, J=12.3Hz, 2H), 1.63 (qd, J=12.1, 4.5Hz, 2H), 1.19 (dd, J=7.6, 6.4Hz, 6H).

[0162] 6-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)hexanoic acid methyl ester (24f)

[0163] Yellow solid, yield 77%. 1H NMR (400MHz, DMSO) δ11.08 (s, 1H), 7.56 (s, 1H), 7.33 (d, J = 8.7Hz, 1H), 7.26 (s, 1H) ),7.07(s,1H),6.89–6.79(m,2H),4.12(s,1H),3.93(d,J=11.4Hz,2H),3.83(s,3 H),3.59(s,3H),3.13–3.06(m,4H),2.59(q,J=7.4Hz,4H),2.32(t,J=7.3Hz,2H), 1.85(d,J=12.8Hz,2H),1.73–1.60(m,4H),1.55(s,2H),1.31(s,2H),1.18(s,3H).

[0164] 7-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)heptanoic acid methyl ester (24 g)

[0165] Yellow solid, yield 79.3%. 1 H NMR (400MHz, DMSO) δ11.01(s,1H),7.52(s,1H),7.30–7.18(m,2H),7.04(d,J=2.3Hz,1H),6.82(d ,J=8.6Hz,1H),6.77(d,J=7.4Hz,1H),3.92(d,J=10.8Hz,2H),3.80(s,3H),3.58(s,3H),3.39(s, 2H),2.92(s,2H),2.58(q,J=7.4Hz,4H),2.30(t,J=7.3Hz,2H),1.85(d,J=12.4Hz,2H),1.63(dd, J=11.8,4.2Hz,2H),1.52(d,J=7.6Hz,2H),1.44(s,2H),1.35–1.25(m,4H),1.19(t,J=7.3Hz,3H).

[0166] 8-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)octanoic acid methyl ester (24h)

[0167] Yellow solid, yield 71.4%. 1H NMR (400MHz, DMSO) δ11.03(s,1H),7.54(d,J=3.4Hz,1H),7.31–7.21(m,2H),7.04(d,J=2.3Hz ,1H),6.81(t,J=8.4Hz,2H),4.17–4.06(m,1H),3.96–3.88(m,2H),3.81(s,3H),3.58(s,3H), 3.38(dd,J=12.0,2.1Hz,2H),2.92(s,4H),2.58(q,J=7.4Hz,2H),2.31(d,J=7.3Hz,2H),1.90 –1.81(m,2H),1.67–1.57(m,2H),1.56–1.43(m,4H),1.31–1.25(m,6H),1.19(t,J=7.4Hz,3H).

[0168] (E)-methyl 3-(4-((4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)phenyl)acrylate (24i)

[0169] Yellow solid, yield 80.6%. 1 H NMR (400MHz, DMSO) δ11.01(s,1H),7.72–7.62(m,3H),7.52(s,1H),7.39(d,J=7.8Hz,2H),7.31 –7.24(m,1H),7.20(s,1H),7.03(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),6.77(d,J=7.6Hz,1H) ,6.62(d,J=16.0Hz,1H),3.92(d,J=10.3Hz,2H),3.80(s,3H),3.73(s,3H),3.56(s,2H),2.93( s,4H),2.58(q,J=7.4Hz,4H),1.85(d,J=12.6Hz,2H),1.70–1.57(m,2H),1.18(t,J=7.6Hz,3H).

[0170] 6-Ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25a)

[0171] Pale yellow solid, yield 31.3%, mp 184-185°C. 1H NMR (400MHz, DMSO) δ11.04(s,1H),10.35(s,1H),8.64(s,1H),7.53(s,1H),7.30(d,J=8.1Hz,1H),7.22(s,1 H),7.04(d,J=2.3Hz,1H),6.81(dd,J=12.8,8.0Hz,2H),4.12(s,1H),3.92(d,J=10.9Hz,2H),3.83(s,3H),3 .57(s,4H),3.40(s,2H),2.90(s,4H),2.62–2.56(m,2H),2.32(t,J=7.4Hz,2H),1.96(q,J=8.7Hz,2H),1.85 (d,J=12.7Hz,2H),1.64(d,J=12.5Hz,2H),1.55–1.46(m,4H),1.25(d,J=13.1Hz,2H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ171.01,170.11,169.39,152.82,152.21,150.57,136.34,135.99,132.10,118.96,112.15,112.00,10 5.12,66.90,56.18,51.47,51.02,48.06,45.77,41.74,32.64,28.88,25.51,25.04,24.69,11.41.HRMS-ESI(m / z)calc.for C 30 H 44 N8O6[M+H] + 613.3771,found 613.3775.HPLC t R =7.19min (98.56% purity).

[0172] 6-Ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25b)

[0173] Pale red solid, yield 40.7%, mp 164-166°C. 1H NMR (400MHz, DMSO) δ11.06(s,1H),10.34(s,1H),8.67(d,J=1.6Hz,1H),7.54(d,J=3.1Hz,1H),7.35–7.23(m,2H), 7.03(d,J=2.3Hz,1H),6.82(t,J=7.9Hz,2H),4.12(d,J=8.2Hz,1H),3.92(d,J=10.4Hz,2H),3.83(s,3H),3.57(s,4 H),3.38(d,J=12.0Hz,2H),2.90(s,2H),2.84(s,2H),2.58(q,J=7.3Hz,2H),2.32(t,J=7.4Hz,2H),1.93(t,J=7.4H z,2H),1.85(d,J=12.3Hz,2H),1.70–1.57(m,2H),1.49(d,J=6.9Hz,4H),1.32–1.23(m,4H),1.19(t,J=7.4Hz,3H). 13 C NMR (101MHz, DMSO) δ171.07,170.12,169.61,152.86,152.22,150.59,136.35,136.04,132.09,118.99,112.21,112.09,105.25,66.9 0,56.21,51.49,51.04,48.07,45.81,41.77,32.73,32.70,32.66,28.99,28.92,25.51,25.21,24.67,11.40.HRMS-ESI(m / z)calc.for C 31 H 46 N8O6[MH] - 625.3438,found 625.3434.HPLC t R =7.59min (95.52% purity).

[0174] 6-Ethyl-3-((4-(4-(3-(hydroxyamino)-3-oxopropyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25c)

[0175] Yellow solid, yield 35.1%, mp 167-168°C. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.44(s,1H),8.75(s,1H),7.53(d,J=3.3Hz,1H),7 .31–7.18(m,2H),7.05(d,J=2.3Hz,1H),6.81(dd,J=15.8,8.0Hz,2H),4.12(h,J=6.4H z,1H),3.93(dd,J=11.5,3.6Hz,2H),3.82(s,3H),3.40(s,2H),2.94(s,4H),2.59(q,J =7.3Hz,4H),2.20(s,2H),1.90–1.81(m,2H),1.68–1.59(m,2H),1.20(t,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO)δ170.11,152.70,152.21,150.61,135.95,132.03,118.53,112.20,112.1 4,105.38,66.90,56.17,53.16,50.61,48.04,32.64,24.67,11.42.HRMS-ESI(m / z)calc.for C 26 H 38 N8O5[M+H] + 543.3049,found 543.3054.HPLC t R =10.97min (99.93% purity).

[0176] 6-Ethyl-3-((4-(4-(4-(hydroxyamino)-4-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25d)

[0177] Yellow solid, yield 30.9%, mp 169-171°C. 1H NMR (400MHz, DMSO) δ11.05(s,1H),10.45(s,1H),8.73(s,1H),7.54(s,1H),7.30(d,J=8. 6Hz,1H),7.23(s,1H),7.05(s,1H),6.82(dd,J=14.0,8.0Hz,2H),4.16–4.07(m,1H),3.9 2(d,J=11.2Hz,2H),3.82(s,3H),3.38(d,J=11.9Hz,2H),3.02(s,4H),2.59(q,J=7.3Hz, 2H), 2.03(t,J=7.2Hz,2H),1.92–1.72(m,4H),1.69–1.57(m,2H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.20,152.68,152.21,150.58,136.19,132.09,118.61,112.13,112.08,1 05.25,66.90,57.03,56.17,52.90,49.86,48.06,32.63,30.39,24.68,11.42.HRMS-ESI(m / z)calc.forC 27 H 40 N8O5[M+H] + 557.3162,found557.3157.HPLC t R =9.18min (99.13% purity).

[0178] 6-Ethyl-3-((4-(4-(5-(hydroxyamino)-5-oxopentyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25e)

[0179] Yellow solid, yield 34%, mp 167-169°C. 1H NMR (400MHz, DMSO) δ11.00(s,1H),10.32(s,1H),8.63(s,1H),7.51(d,J=3.3Hz,1H),7.25(dd,J=8.6,2.3Hz,1H),7.18(d ,J=3.2Hz,1H),7.05(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.75(d,J=7.5Hz,1H),4.18–4.06(m,1H),3.97–3.86(m,2H), 3.81(s,3H),3.41–3.32(m,2H),2.92(s,4H),2.58(q,J=7.4Hz,2H),2.48(s,2H),2.30(t,J=7.1Hz,2H),1.97(t,J=7.2Hz, 2H), 1.85 (dd, J=12.9, 4.2Hz, 2H), 1.68–1.58 (m, 2H), 1.51 (q, J=7.4Hz, 2H), 1.43 (q, J=7.6Hz, 2H), 1.19 (t, J=7.4Hz, 3H). 13 C NMR (101MHz, DMSO) δ170.11,169.20,152.68,152.21,150.58,136.19,132.09,118.61,112.13,112.08, 105.25,66.90,57.03,56.17,52.90,49.86,48.06,32.63,30.39,24.68,11.42.HRMS-ESI(m / z)calc.for C 28 H 42 N8O5[M+H] + 571.3317,found571.3312.HPLC t R =7.12min (95.58% purity).

[0180] 6-Ethyl-3-((4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25f)

[0181] Yellow solid, yield 37.2%, mp 169-171°C. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.35(s,1H),8.69(s,1H),7.54(d,J=3.2Hz,1H),7.29–7.22(m,2H),7 .03(d,J=2.4Hz,1H),6.81(t,J=7.4Hz,2H),4.12(d,J=9.5Hz,1H),3.92(d,J=11.4Hz,2H),3.80(s,3H),2. 91(s,4H),2.59(t,J=7.3Hz,2H),2.28(t,J=7.3Hz,2H),1.95(t,J=7.3Hz,2H),1.85(d,J=12.6Hz,2H),1.6 8–1.59(m,2H),1.51(q,J=7.4Hz,2H),1.44(q,J=7.0Hz,2H),1.26(q,J=7.8Hz,2H),1.19(t,J=7.4Hz,3H). 13 C NMR (101MHz, DMSO) δ170.12,169.50,152.70,152.21,150.61,136.61,135.81,131.98,118.44,112.22,112.13,105.42 ,66.90,58.39,56.17,53.62,50.95,48.04,32.72,32.65,27.03,26.51,25.58,24.67,11.42.HRMS-ESI(m / z)calc.for C 29 H 44 N8O5[M+H] + 585.3472,found585.3467.HPLC t R =8.60min (99.22% purity).

[0182] 6-ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25 g)

[0183] Yellow solid, yield 31.9%, mp 199-201°C. 1H NMR (400MHz, DMSO) δ11.06(s,1H),10.36(s,1H),8.67(s,1H),7.54(d,J=3.2Hz,1H),7.31(d,J=8.6Hz,1H),7.2 3(d,J=3.3Hz,1H),7.05(d,J=2.3Hz,1H),6.83(dd,J=17.4,8.0Hz,2H),4.11(p,J=5.3Hz,1H),3.93(dd,J=11.4, 3.7Hz,2H),3.82(s,3H),3.38(d,J=10.6Hz,2H),3.02(s,4H),2.59(q,J=7.3Hz,2H),1.95(t,J=7.3Hz,2H),1.86 (dd,J=12.9,4.1Hz,2H),1.71–1.54(m,4H),1.50(t,J=7.1Hz,2H),1.28(d,J=6.9Hz,4H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.53,152.68,152.21,150.57,136.43,132.14,118.72,112.19,112. 01,105.19,66.89,56.18,48.06,32.63,28.72,26.65,25.42,24.69,11.42.HRMS-ESI(m / z)calc.for C 30 H 46 N8O5[M+H] + 599.3626,found599.3621.HPLC t R =8.60min (99.22% purity).

[0184] 6-Ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25h)

[0185] Yellow solid, yield 39.6%, mp 205-207°C. 1H NMR (400MHz, DMSO) δ11.03(s,1H),10.34(s,1H),8.67(s,1H),7.54(d,J=3.1Hz,1H),7.31–7.21(m,2H) ,7.03(d,J=2.3Hz,1H),6.81(t,J=7.7Hz,2H),4.18–4.07(m,1H),3.92(d,J=11.4Hz,2H),3.81(s,3H),3 .39(s,2H),2.93(s,4H),2.61–2.57(m,2H),2.34(s,2H),1.94(t,J=7.3Hz,2H),1.89–1.82(m,2H),1.6 9–1.60(m,2H),1.54–1.44(m,4H),1.25(d,J=12.5Hz,6H),1.19(t,J=7.3Hz,3H),1.13(t,J=7.3Hz,2H). 13 C NMR (101MHz, DMSO) δ170.10,169.56,152.68,152.21,150.56,132.14,118.73,112.19,111.99,105.16, 66.89,56.18,49.06,48.06,32.70,32.64,28.87,26.77,25.49,24.69,11.42.HRMS-ESI(m / z)calc.for C 31 H 48 N8O5[M+H] + 613.3767,found613.3762.HPLC t R =7.66min (99.55% purity).

[0186] (E)-6-ethyl-3-((4-(4-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25i)

[0187] Yellow solid, yield 38.1%, mp 200-201°C. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.79(s,1H),9.04(s,1H),7.53(d,J=6.3Hz,3H),7.45(d,J=15.7Hz,1 H),7.37(d,J=7.7Hz,2H),7.31–7.20(m,2H),7.03(d,J=2.2Hz,1H),6.81(dd,J=13.2,8.0Hz,2H),6.47(d, J=15.8Hz,1H),4.19–4.05(m,1H),3.92(dd,J=11.7,3.6Hz,2H),3.80(s,3H),3.54(s,2H),3.39(s,2H),2 .94(s,4H),2.59(q,J=7.2Hz,2H),1.85(dd,J=12.6,4.1Hz,2H),1.68–1.60(m,2H),1.19(t,J=7.2Hz,3H). 13 C NMR (101MHz, DMSO) δ170.12,163.29,152.73,152.21,150.61,138.61,136.50,135.90,134.07,132.01,129.93,127.84,119 .15,118.55,112.17,112.12,105.34,66.90,62.24,56.16,53.37,50.93,48.04,32.64,24.67,11.42.HRMS-ESI(m / z)calc.C 33 H 42 N8O5[M+H] + 631.3312,found 631.3306.HPLC t R =6.43min (95.75% purity).

[0188] (E)-6-ethyl-3-((4-(4-(4-((4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)amino)-2-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25j)

[0189] Pale yellow solid, yield 37.9%. 1H NMR(400MHz,DMSO)δ11.05(s,1H),10.84(s,1H),9.05(s,1H),8.47(s,1H),7.63–7.39(m,4H), 7.36–7.17(m,4H),7.05(s,1H),6.83(d,J=8.6Hz,2H),6.49(d,J=15.8Hz,1H),4.29(d,J=5.8Hz ,2H),4.12(s,1H),3.98–3.88(m,2H),3.82(s,3H),3.03(s,4H),2.76(s,3H),2.61–2.56(m,2H ), 2.23 (t, J = 7.4Hz, 2H), 1.85 (d, J = 15.5Hz, 4H), 1.65 (d, J = 12.4Hz, 2H), 1.19 (t, J = 7.3Hz, 3H). 13 C NMR (101MHz, DMSO) δ172.28,170.12,163.23,152.69,152.22,150.59,141.66,138.45,136.13,133.86,132.09,128.19,127.92,119 .21,118.58,112.14,105.31,66.90,57.21,56.19,52.97,49.96,48.06,42.30,33.42,32.63,24.69,11.42.HRMS-ESI(m / z)calc.for C 37 H 49 N9O6[MH] - 713.4716,found 713.4712.HPLC t R =2.78min (96.57% purity).

[0190] (E)-6-ethyl-3-((4-(4-(2-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25k)

[0191] Pale yellow solid, yield 34.5%. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.67(s,1H),8.98(s,1H),7.52(dd,J=12.1,5.7Hz,3H),7.41(d,J=15.7Hz,1H ),7.30–7.20(m,2H),7.05(d,J=2.3Hz,1H),7.00(d,J=8.3Hz,2H),6.81(dd,J=14.3,8.0Hz,2H),6.32(d,J=15.8H z,1H),4.16(s,2H),4.11(t,J=8.4Hz,1H),3.92(d,J=11.5Hz,2H),3.81(s,3H),3.39(s,2H),2.95(s,4H),2.79(s ,2H),2.67(s,2H),2.58(d,J=7.4Hz,2H),1.85(dd,J=12.8,4.1Hz,2H),1.70–1.57(m,2H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.12,163.66,159.97,152.71,152.21,150.62,138.49,136.43,135.93,132.01,129.52,127.93,118.52,117. 04,115.40,112.23,112.14,105.42,66.90,65.96,56.98,56.18,53.82,50.74,48.04,32.65,24.67,11.42.HRMS-ESI(m / z)calc.for C 34 H 44 N8O6[M+H] + 661.3416,found 661.3411.HPLC t R =11.59min(99.81%purity).

[0192] 2-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)-N-hydroxypyrimidine-5-carboxamide (251)

[0193] Pale yellow solid, yield 33.1%. 1H NMR (400MHz, DMSO) δ11.46(s,1H),8.79(s,2H),7.63(s,2H),7.22(s,1H),6.97(d,J=7.4Hz,1H),4.13(s,1H),4.05–3.88(m ,7H),3.60(s,4H),2.62(q,J=7.3Hz,2H),1.86(dd,J=12.9,3.5Hz,2H),1.67(qd,J=12.1,4.5Hz,3H),1.20(t,J=7.4Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,163.63,160.13,152.69,152.21,150.58,138.48,136.27,132.10,129.52,127.83,118.67,11 6.99,115.34,112.17,112.08,105.26,67.73,66.90,56.18,48.05,32.64,26.72,24.68,11.42.HRMS-ESI(m / z)calc.forC 36 H 48 N8O6[M+H] + 689.3725,found689.3722.HPLC t R =6.40min (96.92% purity).

[0194] 6-Ethyl-3-((4-(4-(2-((4-(hydroxyamino)-4-oxobutyl)(methyl)amino)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25m)

[0195] Pale yellow solid, yield 31.7%. 1H NMR (400MHz, DMSO) δ11.02(s,1H),10.35(s,1H),8.65(s,1H),7.53(s,1H),7.29–7.20(m,2H),7. 04(d,J=2.3Hz,1H),6.80(t,J=8.7Hz,2H),4.11(s,1H),3.91(t,J=6.8Hz,2H),3.81(s,3H),3.39 (s,2H),2.91(s,4H),2.59(t,J=7.3Hz,2H),2.43(s,4H),2.28(t,J=7.2Hz,2H),2.15(s,2H),1.9 7(q,J=8.6Hz,2H),1.90–1.81(m,2H),1.63(q,J=6.8Hz,4H),1.23(s,2H),1.19(t,J=7.3Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.35,152.71,152.21,150.60,136.40,135.93,132.01,118.49,112.19,112.15,105.39,66 .90,60.23,56.19,53.62,50.70,48.04,45.96,32.65,32.37,24.68,23.13,21.24,14.56,11.43.HRMS-ESI(m / z)calc.for C 31 H 49 N9O5[M+H] + 614.3756,found 614.3751.HPLC t R =6.88min (97.52% purity).

[0196] 6-Ethyl-3-((4-(4-(2-((4-(hydroxycarbamoyl)benzyl)(methyl)amino)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25n)

[0197] Pale yellow solid, yield 35.2%. 1H NMR (400MHz, DMSO) δ11.22(s,1H),11.05(s,1H),7.73(d,J=7.8Hz,2H),7.56(d,J=3.3Hz,1H),7. 42(d,J=7.6Hz,2H),7.29(d,J=9.3Hz,2H),7.04(d,J=2.4Hz,1H),6.87–6.77(m,2H),4.18–4.08(m ,1H),3.97–3.89(m,2H),3.81(s,3H),3.63(s,2H),3.05(q,J=7.5Hz,4H),2.96(s,2H),2.58(q,J =7.4Hz,5H),2.20(s,2H),1.89–1.81(m,2H),1.63(qd,J=12.4,4.5Hz,2H),1.20(d,J=3.7Hz,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.00,152.69,152.21,150.60,136.32,135.96,132.02,118.51,112.13,105.34 ,66.90,56.17,53.33,50.63,48.05,45.90,32.64,30.09,24.68,14.56,11.43,9.21.HRMS-ESI(m / z)calc.for C 34 H 47 N9O5[M+H] + 662.3451,found 662.3746.HPLC t R =8.41min (97.95% purity).

[0198] (E)-6-ethyl-3-((4-(4-(2-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)(methyl)amino)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25o)

[0199] Pale yellow solid, yield 37.2%. 1H NMR (400MHz, DMSO) δ11.04(s,1H),10.76(s,1H),7.56–7.51(m,2H),7.45(d,J=15.8Hz,1H),7.38(s,1 H),7.26(dd,J=14.9,6.0Hz,2H),7.05(d,J=2.4Hz,1H),6.82(t,J=8.6Hz,2H),6.46(d,J=15.8Hz,1H) ,4.11(s,1H),3.96–3.89(m,2H),3.81(s,3H),3.51(s,2H),3.06(d,J=7.4Hz,4H),2.96(s,2H),2.58( q,J=7.3Hz,5H),2.22(s,2H),1.85(dd,J=13.1,4.1Hz,2H),1.63(qd,J=12.1,4.5Hz,2H),1.19(s,3H). 13 C NMR (101MHz, DMSO) δ170.11,169.00,152.69,152.21,150.60,136.32,135.96,132.02,118.51,112.13,105.34 ,66.90,56.17,53.33,50.63,48.05,45.90,32.64,30.09,24.68,14.56,11.43,9.21.HRMS-ESI(m / z)calc.for C 36 H 49 N9O5[M+H] + 688.3898,found688.3895.HPLC t R =9.40min (97.47% purity).

[0200] 2-((3-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)propyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (25p)

[0201] Pale yellow solid, yield 37.2%. 1H NMR (400MHz, DMSO) δ11.06(s,1H),9.00(s,1H),8.68(s,2H),7.54(d,J=3.4Hz,1H),7.30(d,J=8.4H z,1H),7.24(d,J=3.1Hz,1H),7.05(d,J=2.3Hz,1H),6.83(dd,J=11.5,8.1Hz,2H),4.13(s,1H),3.92 (dd,J=11.2,3.6Hz,2H),3.82(s,3H),3.69(d,J=7.5Hz,2H),3.42–3.36(m,2H),3.16(s,3H),2.96(s ,2H),2.59(q,J=7.4Hz,4H),1.88–1.82(m,2H),1.64(qd,J=12.2,4.3Hz,2H),1.19(t,J=7.4Hz,3H). 13 CNMR(101MHz,DMSO)δ170.11,169.00,152.69,152.21,150.60,136.32,135.96,132.02,118.51,112.13,105.3 4,66.90,56.17,53.33,50.63,48.05,45.90,32.64,30.09,24.68,14.56,11.43,9.21.HRMS-ESI(m / z)calc.for C 32 H 45 N 11 O5[M+H] + 664.3651,found664.3646.HPLC t R =8.77min (96.88% purity).

[0202] 2-(4-((4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25q)

[0203] Pale yellow solid, yield 35.3%. 1H NMR (400MHz, DMSO) δ11.05(d,J=10.9Hz,2H),8.99(s,1H),8.66(s,2H),7.54(d,J=3.3Hz,1H),7.30(d,J=8.5 Hz,1H),7.26–7.21(m,1H),7.04(d,J=2.4Hz,1H),6.82(dd,J=13.4,8.0Hz,2H),4.71(d,J=12.8Hz,2H),4.16– 4.06(m,1H),3.96–3.89(m,2H),3.82(s,3H),3.42–3.35(m,2H),2.94(d,J=12.0Hz,6H),2.59(q,J=7.4Hz,2H) ,2.19(s,2H),1.86(dd,J=12.9,3.9Hz,4H),1.64(qd,J=12.1,4.3Hz,2H),1.19(t,J=7.3Hz,3H),1.06(s,2H). 13 C NMR (101MHz, DMSO) δ170.11,169.00,152.69,152.21,150.60,136.32,135.96,132.02,118.51,112.13,105.34 ,66.90,56.17,53.33,50.63,48.05,45.90,32.64,30.09,24.68,14.56,11.43,9.21.HRMS-ESI(m / z)calc.forC 34 H 47 N 11 O5[M+H] + 690.3802,found 690.3798.HPLC t R =9.87min (95.99% purity).

[0204] Experimental Example 1: Inhibitory activity of target compounds on HDACs

[0205] 1. Materials:

[0206] HDACs enzyme (HeLa cell nuclear extract); Boc-Lys(Ac)-AMC substrate; trypsin; EDTA; LBH-589 (10 mM, dissolved in dimethyl sulfoxide); 96-well all-black ELISA plate; glycerol; ultrapure water

[0207] 2. Methods:

[0208] Buffer preparation:

[0209] The formula is 15 mM Tris-HCl (pH=8.0), 250 μM EDTA, 250 mM NaCl, and 10% glycerol.

[0210] Preparation of Trypsin solution:

[0211] The formula is 10 mg / mL trypsin, Buffer, 2 μM LBH589.

[0212] Preparation of substrate solution:

[0213] Dissolve the substrate in DMSO to prepare a 30 mM stock solution, then dilute to 300 μM with buffer.

[0214] The content of DMSO is about 1%.

[0215] Dilution of enzyme solution:

[0216] Dilute the enzyme solution with buffer at a ratio of 1:80.

[0217] Preparation of compound solution:

[0218] Dilute the compounds (test compounds and positive control drug SAHA) to 5× final concentration with buffer

[0219] Preparation and determination of 100% and blank:

[0220] 50μL buffer was mixed with 10μL enzyme solution. After 5 minutes, 40μL substrate was added and reacted at 37℃ for 0.5 hour. Then 100μL Trypsin solution was added to terminate the reaction. After reacting at 37℃ for 20 minutes, the fluorescence intensity was measured at (390nm / 460nm) to obtain 100% absorption. After 40μL substrate was added to 60μL buffer and reacted at 37℃ for 0.5 hour, 100μL Trypsin solution was added and reacted at 37℃ for 20 minutes to obtain blank absorption.

[0221] Determination of the inhibition of HDACs activity by compounds:

[0222] 50 μL of drug-containing buffer was mixed with 10 μL of enzyme solution and pre-incubated for 5 minutes. After adding 40 μL of substrate, the reaction was allowed to proceed at 37°C for 0.5 hours. Then, 100 μL of Trypsin solution was added to terminate the reaction. After reacting at 37°C for 20 minutes, the fluorescence intensity was measured at (390 nm / 460 nm). The inhibition rate and IC were calculated using GraphPad Prism software and the formula 50 Table 1. HDACs inhibitory activity of target compounds

[0223]

[0224]

[0225] a A,IC 50 Greater than 5 μM; B, IC 50 Between 1μM and 5μM; C, IC 50 Between 0.2μM and 1μM; D, IC 50 Less than 0.2μM

[0226] It can be seen from Table 1 that compounds 11j, 25b, 25h-k and 25o-q have strong inhibitory activity against HDAC enzymes, which is comparable to the positive control drug SAHA. The remaining compounds also show good inhibitory activity against HDAC enzymes.

[0227] Experimental Example 2: Inhibitory activity of target compounds on FLT3

[0228] 1. Materials:

[0229] FLT3 enzyme; 96-well all-black ELISA plate; 40 mM Tris buffer (pH=7.4); MgCl2; bovine serum albumin (BSA); dithiothreitol (DTT); adenosine triphosphate (ATP); FLT3 ligand; positive control drug gilteritinib.

[0230] 2. Methods:

[0231] The target compound (5 μL) at the specified concentration was added to a 50 μL reaction system containing 40 mM Tris (pH = 7.4), 10 mM MgCl2, 0.1 mg / ml BSA, 1 mM DTT, 10 μM ATP, FLT3 enzyme and enzyme substrate, and incubated at 30 ° C for 40 minutes. Then, the assay was performed according to the protocol of the Kinase-glo Plus Fluorescent Kinase Assay Kit. The kinase inhibitory activity is proportional to the remaining amount of ATP. The fluorescence intensity was detected with a microplate reader, and the inhibitory activity was calculated using GraphPad Prism software (Table 2).

[0232] Table 2. FLT3 inhibitory activity of target compounds

[0233]

[0234]

[0235] a A,IC 50 Greater than 50nM; B, IC 50Between 5nM and 50nM; C, IC 50 Between 2nM and 5nM; D, IC 50 Less than 2nM

[0236] It can be seen from Table 2 that compounds 25e-h and 25k-q have the strongest inhibitory effect on FLT3 enzyme activity, which is comparable to the positive control drug gilteritinib. In addition, compounds 11c, 25a-d, and 25j also have strong inhibitory activity on FLT3, with IC 50 Values ​​ranged from 2 to 5 nM.

[0237] Experimental Example 3: Antiproliferative Activity Experiment of Target Compound

[0238] 1. Materials:

[0239] MV4-11, MOLM-13, KG-1, THP-1, Jurkat, K562 cell lines, 10% fetal bovine serum (Hyclone, USA), 2.5 g·L -1 Trypsin (Gibco, USA), CCK-8, modified RPMI1640 medium, DMEM medium (Hyclone, USA), positive control drugs SAHA, GSK3685032, 96-well plates.

[0240] 2. Methods:

[0241] Culture cells routinely and collect logarithmically growing cells for experiments; dilute logarithmically growing solid tumor cells to 4×10 3 pc·mL -1 (Hematologic tumor cells were diluted to 1×10 4 pc·mL -1 ) were inoculated into a 96-well plate (100 μL was added to each well), and the wells without cells were used as blank wells, and then placed in a constant temperature incubator (37°C, 5% carbon dioxide) for 8 hours; the target compound solution and positive drug solution prepared with culture medium were added, and the wells without drugs were used as 100% wells. After culturing in a constant temperature incubator (37°C, 5% carbon dioxide) for 72 hours, 20 μL CCK-8 was added, and the absorbance value of each well was measured at a wavelength of 450 nm using an enzyme reader after three hours, and the inhibition rate and IC 50 Values ​​(Table 3).

[0242] Table 3. Antiproliferative activity of selected compounds

[0243]

[0244]

[0245] aA,IC 50 Greater than 20 μM; B, IC 50 Between 2μM and 20μM; C, IC 50 Between 0.2μM and 2μM; D, IC 50 Less than 0.2μM

[0246] It can be seen from Table 3 that the compounds with good inhibitory activity against both FLT3 and HDAC enzymes are 25g, 25h, 25j, 25k and 25o-q, which can effectively inhibit the proliferation of acute myeloid leukemia cells with FLT3-ITD mutations, such as MV4-11 and MOLM-13 cells, and the effect is comparable to that of gilteritinib and stronger than SAHA. In addition, compounds 25h, 25k and 25o-q also show strong anti-proliferative activity against other types of acute leukemia (KG-1, THP-1 and Jurakat) and chronic myeloid leukemia cell lines, and the effect is better than that of gilteritinib and comparable to that of SAHA. Therefore, these compounds have further research value.

[0247] Experimental Example 4: Study on the therapeutic effect of compounds on human acute myeloid leukemia cells

[0248] 1. Materials:

[0249] Cell line: Human acute myeloid leukemia cells (MOLM-13).

[0250] Experimental animals: Six-week-old BALB / c female nude mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.

[0251] 2. Methods:

[0252] About 2×10 7 MOLM-13 cells were resuspended in 50 μL serum-free 1640 medium and 50 μL Matrigel and implanted subcutaneously in the right axillary region of BALB / c nude mice. 3 At 14:00, oral administration was started for 9 consecutive days. The treatment group dose was 10 mg / kg / d (25p) and 10 mg / kg / d + 30 mg / kg / d (gilteritinib + SAHA), and the control group was given an equal amount of normal saline. Tumor size and mouse body weight were measured every day during the treatment period. Tumor volume calculation formula: V (mm 3 )=Length(mm)×Width 2 (mm) / 2; Tumor growth inhibition value (TGI) calculation formula: TGI=(1-average tumor weight of treatment group / average tumor weight of control group)×100%.

[0253] 3. Experimental results:

[0254] The results are as follows Figure 1As shown, the tumor inhibition rate of the target compound 25p is as high as 64%, and the tumor inhibition effect is better than that of the combined treatment group of the positive control drug gilteritinib and SAHA.

Claims

1. A FLT3-HDAC dual-target inhibitor or a pharmaceutically acceptable salt or a stereoisomer thereof, characterized in that: The FLT3-HDAC dual-target inhibitor has a structure shown in Formula I: In formula I, R1 is hydroxyl or 2-aminophenyl, X is an aromatic heterocycle or one of the following linking groups: A is one of the following structures: R2 is a hydrogen atom or one of the following structures 2. The FLT3-HDAC dual-target inhibitor according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: In the FLT3-HDAC dual-target inhibitor formula I, R1 is hydroxyl or 2-aminophenyl; X is one of the following linking groups: A is piperazinyl or piperidin-4-ylpiperazinyl; R2 is tetrahydropyranylamino.

3. The FLT3-HDAC dual-target inhibitor according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The FLT3-HDAC dual-target inhibitor is selected from one of the following compounds: 6-ethyl-3-((4-(4-(4-(5-(hydroxyamino)-5-oxopentanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11a); 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxohexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11b); 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxoheptanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11c); 6-ethyl-3-((4-(4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11d); 6-ethyl-3-((4-(4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11e); 6-ethyl-3-((4-(4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11f); 6-ethyl-3-((4-(4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11 g); 6-ethyl-3-((4-(4-(4-(4-(hydroxycarbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11h); (E)-6-ethyl-3-((4-(4-(4-(4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11i); (E)-6-ethyl-3-((4-(4-(4-(4-((4-(3-hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)carbamoyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (11j); 2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)N-hydroxy-pyrimidine-5-carboxamide (14); 3-((4-(4-(4-(4-((2-aminophenyl)benzyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (17a); N-(2-aminophenyl)-2-(4-(1-(4-((3-carbamoyl-5-ethyl-6((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pyrimidine-5-carboxamide (17b); 6-ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25a); 6-ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctanoyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25b); 6-ethyl-3-((4-(4-(3-(hydroxyamino)-3-oxopropyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25c); 6-ethyl-3-((4-(4-(4-(hydroxyamino)-4-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25d); 6-ethyl-3-((4-(4-(5-(hydroxyamino)-5-oxopentyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25e); 6-ethyl-3-((4-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25f); 6-ethyl-3-((4-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25 g); 6-ethyl-3-((4-(4-(8-(hydroxyamino)-8-oxooctyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25h); (E)-6-ethyl-3-((4-(4-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25i); (E)-6-ethyl-3-((4-(4-(4-((4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)benzyl)amino)-2-oxobutyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25j); (E)-6-ethyl-3-((4-(4-(2-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)ethyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25k); (E)-6-ethyl-3-((4-(4-(3-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)propyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (251); (E)-6-ethyl-3-((4-(4-(4-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenoxy)butyl)piperazin-1-yl)-3-methoxyphenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (25m); 2-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25n); 2-((3-(4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)propyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (25o); 2-(4-((4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25p); 2-(4-((4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide (25q).

4. The method for preparing the FLT3-HDAC dual-target inhibitor according to claim 1, comprising the following steps: Compound 1 reacts with 4-aminotetrahydropyran to obtain compound 2. Meanwhile, compounds 3 and 18 react with 1-fluoro-2-methoxy-4-nitrobenzene to obtain compounds 4 and 19. Compounds 4 and 19 are further reduced to compounds 5 and 20. Compounds 2 and 5 or 20 are subjected to Buchwald-Hartwig coupling reaction to obtain compounds 6 and 21. Compounds 6 and 21 are subjected to oxidation reaction and removal of Boc protecting groups to obtain compounds 8 and 23. Compounds 8 and 23 are further reacted with 9a-u, 12, 15a and 15b to generate 10a-j, 13, 16a, 16b and 24a-q, wherein the ester groups of compounds 10a-j and 24a-q can be converted into hydroxamic acid groups in an alkaline hydroxylamine solution to obtain target compounds 11a-j and 25a-q. Compounds 13, 16a and 16b are subjected to removal of protecting groups in an acidic environment to obtain target compounds 14, 17a and 17b.

5. The method for preparing the FLT3-HDAC dual-target inhibitor according to claim 1, wherein the method comprises any one of the following methods: The preparation methods of compounds 11a-11j are as follows: The preparation methods of compounds 14, 17a and 17b are as follows: The preparation method of compound 25a-q is as follows:

6. The preparation method according to claim 5, characterized in that: During the preparation of compounds 11a-11j, Reagents and conditions: (a) 4-aminotetrahydropyran, triethylamine, acetonitrile, 80°C, 6 hours; (b) 1-fluoro-2-methoxy-4-nitrobenzene, potassium carbonate, N,N-dimethylformamide, 100°C, 8 hours; (c) 10% Pd / C, tetrahydrofuran, room temperature, 24 hours; (d) palladium acetate, 4,5-bis(diphenylphosphino-9,9-dimethylxanthene), cesium carbonate, 1,4-dioxane, 100°C, 12 hours; (e) 30% hydrogen peroxide, potassium carbonate, dimethyl sulfoxide, room temperature, 12 hours; (f) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (g) 1) 9a-d, EDCI, DMAP, room temperature, 8 hours; 2) 9e-j, potassium carbonate, acetonitrile, 80°C, 6 hours; (h) NH2OH, KOH, anhydrous methanol, room temperature, 4 hours.

7. The preparation method according to claim 5, characterized in that: During the preparation of compounds 14, 17a, and 17b, Reagents and conditions: (a) 12, triethylamine, acetonitrile, 80°C, 8 hours; (b) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (c) 15a and 15b, triethylamine, acetonitrile, 80°C, 8 hours; (d) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours.

8. The preparation method according to claim 5, characterized in that: During the preparation of compound 25a-q, Reagents and conditions: (a) 1-fluoro-2-methoxy-4-nitrobenzene, potassium carbonate, N,N-dimethylformamide, 100°C, 8 hours; (b) 10% Pd / C, tetrahydrofuran, room temperature, 24 hours; (c) 2, palladium acetate, 4,5-bis(diphenylphosphino-9,9-dimethylxanthene), cesium carbonate, 1,4-dioxane, 100°C, 12 hours; (d) 30% hydrogen peroxide, potassium carbonate, dimethyl sulfoxide, room temperature, 12 hours; (e) 4M hydrochloric acid / ethyl acetate, room temperature, 12 hours; (f) 1) 9c and 9d, EDCI, DMAP, room temperature, 8 hours; 2) 9e-u, potassium carbonate, acetonitrile, 80°C, 6 hours; (g) NH2OH, KOH, anhydrous methanol, room temperature, 4 hours.

9. A pharmaceutical composition comprising a therapeutically effective amount of the FLT3-HDAC dual-target inhibitor according to claim 1 and a pharmaceutically acceptable excipient.

10. Use of the FLT3-HDAC dual-target inhibitor of claim 1 in the preparation of a drug for treating a target-related tumor disease, wherein the tumor is lung cancer, liver cancer, non-small cell lung cancer, colon cancer, skin cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, lymphoma, esophageal cancer, gastrointestinal cancer, nasopharyngeal carcinoma, leukemia, or myeloma.

Images